The Division of Microbiology provides diagnostic support to all divisions at NERPRC, especially to the following divisions Primate Medicine, Immunology, Pathology, Cardiovascular Medicine and Behavior Biology. In addition we also provide support to the visiting scientists and collaborative scientists using the facilities of our center. Diagnostic Virology The annexed table provides in a tabular form the number of tissues processed for virus isolation. The following viruses were isolated Type D retrovirus, foamy and adeno from macaques. As in the past year we were fortunate that no B virus was isolated from 50 scratch samples of employees. Again as in the recent past we had no spontaneous cases of SIV in our macaque colony. We had considerably more SIV samples for test using the antigen capture test - 8004 versus 6903 in the previous year. Routine screening for Herpes B, SIV STLV-1, TyD were done by Elisa test. We had tested more samples (1279) than the previous year (809) total number of samples tested were 1279. Screening for foamy antibodies were done by immunofluorescence tests. Animals used for virus inoculation studies were screened for foamy virus antibodies by this procedure for selecting foamy negative animals. Serum neutralizing tests were done by using MTT assays in 988 wells. Virus titration for SIV and HIV-1 experiments were done in CEMx174 on 380 plates, involving 8166 wells which is an increase from last year. Measurements for determining the level of virus in infected lymphocytes (virus load) were done in limbro. Colony born owl monkeys were vaccinated with herpes T and h. simplex vaccines. The samples submitted for bacterial analysis are provided in tabular form. No unusual agents were isolated. There was an increase in the number of culture samples this year 1134 as opposed to 826 last year. Diagnostic Bacteriology There were 1134 animal and tissue samples submitted for testing this year. These samples were form the Division of Primate Medicine, Comparative Pathology, Behavioral Biology and Cardiovascular Medicine. Quality Control for Central Supply was monitored weekly as was QC for the virus room. Samples were also tested from environmental sources, water bottles, reagents, buffers and laboratory solutions. This year we have been testing for Clostridium by using Clostridium cytotoxic assay. There were 2475 bacterial and fungal isolations made from a total of 1134 samples submitted. This was an increase from the previous years (2120). The annexed tables provide a breakdown of the organisms isolated from our center. The most frequently isolated organisms was E. coli 455 isolations a 18.38% rate. The pathogenic E.coli strain was much less, with 40 positive isolations, Klebsiella pneumonia was still common in our animal colony with 228 isolations a 9.21% rate. The pathogenic strain of staphaureus was less frequent with 124 positive identifications a 5.01% rate. Next was, streptococcus (Alpha hemolytic) 258 isolations a 10.42%. 1995 Summary Report M.D. Daniel - Rm 204 Tissue Samples lymphs sera stored sera tested oral swab eye swab fecal swab k.s. lesion brain kidney pancrease tumor ____________________________________________________________________________ Mm 806 1205 33 20 17 4 - 2 1 4 - Mf 16 17 - 8 5 - - - - - 1 Mn 43 53 - - - - - - - - - Cpz 88 66 - - - - - - - - - Human 8 4 - - - - 5 - - - - ____________________________________________________________________________ B-virus Samples 50 TyD virus checks 77 Foamy virus iso. attempts 11 SIV40 iso. attempts 11 Adeno virus iso. attempts 12 AC Samples 8,004 MTT/Serum Neuts 988 wells (13 plates) ELISA 1,279 (20 plates) Virus Load 8,166 (380 plates) SAMPLE SUBMITTED FOR BACTERIOLGIC ANALYSIS FOR 1995 Callithrix jacchus 86 Canines 141 Homo sapiens 6 Macaca cyclopis 3 Macaca fascicularis 39 Macaca mulatta 542 Macaca nemistrina 12 Saguinus oepdipus 189 Saimiri sciureus 57 Canines 2 Gerbil 1 Homo sapiens 6 Mice 3 Miscellaneous Cell culture sample 26 Water sample 1 Culture medius sample 20 TOTAL 1134 BACTERIAL AND FUNGAL ISOLATES (TOTAL=2475) ORGANISM J00 PERCENT Acinetobacter anitratus 10 0.40 Acinetobacter lwoffi 12 0.48 Actinomyces viscosus 1 0.04 Alcaligenes species 2 0.08 Bacillus species 27 1.09 Bacteroids fragila group 4 0.16 Bacteroids sp. 1 0.04 Bordetella bronchiseptica 2 0.08 Branhamella catarrhalis 5 0.20 CDC group EF-4 1 0.04 Campylobacter jejuni 126 5.09 Candida albican 3 0.12 Citrobacter diversus 1 0.04 Citrobacter freundii 15 0.61 Clostridium difficile 4 0.16 Corynebacteria species 194 7.84 Cryptococcus terreus 1 0.04 Enterobacter aerogenes 1 0.04 Enterobacter agglomerans 22 0.89 Enterobacter cloacae 30 1.21 Enterobacter sakazakii 1 0.04 Enterobacter taylorae 1 0.04 Enterococcus 259 10.46 Escherichia coli 455 18.38 Escherichia coli (Beta hemolytic) 40 1.62 Escherichia vulneris 1 0.04 Fungi (yeast) 19 0.77 Fusobacterium nucleatum 1 0.04 Fusobacterium species 3 0.12 Haemophilus parainfluenzae 4 0.16 Haemophilus species 14 0.57 Halfnia alvei 1 0.04 Klebsiella oxytoca 3 0.12 Klebsiella ozaenae 1 0.04 Klebsiella pneumoniae 228 9.21 Micrococcus luteus 2 0.08 Morganella morganii 20 0.81 Moraxella species 1 0.04 Neisseria species 3 0.12 Other bacteroides species 3 0.12 Peptostreptococcus indole 1 0.04 Proteus mirabilis 108 4.36 Proteus vulgaris 4 0.16 Providencia alcalifaciens 1 0.05 Providencia rettgeri 8 0.32 Pseudomonas aeruginosa 64 2.59 Pseudomonas fluor/putida 1 0.04 Pseudomonas maltophilia 1 0.04 Serratia marcescens 5 0.20 Shigella flexneri type 4 10 0.40 ORGANISM J00 PERCENT Staphylococcus aureus 121 5.01 Staphycoccus coag. negative 359 14.51 Staphycoccus hyicus 1 0.01 Streptococcus (Alpha hemolytic) 258 10.42 Streptococcus not Group A 6 0.24 Streptococcus, Group A 2 0.08